摘要
目的:研究NF-κB对人非霍奇金淋巴瘤(NHL)HIF-1α-VEGF途径的调控及其机制。方法:应用Echinomycin(EC)处理人NHL细胞,将HIF-1α反义质粒转染至肿瘤细胞中,采用蛋白质印迹法检测经两种方法处理后各细胞系中VEGF表达水平。以NF-κB特异性抑制剂quinazoline(QNZ)及Bay11-7082预处理人NHL细胞,检测各细胞系中HIF-1α蛋白的表达水平,同时采用实时定量PCR方法检测HIF-1αmRNA变化。应用QNZ处理NHL细胞后检测HIF-1α下游调控靶点VEGF的蛋白表达水平。结果:通过应用HIF-1α特异性抑制剂和转染反义质粒两种方法均能够抑制HIF-1α,明显下调了VEGF蛋白表达水平,与对照组相比差异均有统计学意义,P<0.05。NF-κB抑制剂QNZ及Bay11-7082能够降低NHL细胞HIF-1α蛋白及基因水平的表达,同时下调其下游调控靶点VEGF蛋白的表达,P<0.05。结论:抑制NF-κB可阻断人NHL细胞HIF-1α-VEGF通路,其机制可能与NF-κB调控HIF-1α的基因与蛋白表达有关。
OBJECTIVE：To investigate the role of NF-κB in regulation of HIF-1α expression of human non-Hodgkin lymphoma （NHL） cells in normoxic condition,as well as to explore the mechanism mediating this effect. METHODS： Ex pression levels of VEGF protein in three human non-Hodgkin lymphoma cell lines, Namalwa, Ramos, and Raji cell were determined by Western blot after treatment with EC,a HIF-1α antagonist or with pSuper HIF-1α/siRNA transfection. Cells were pre-treated by NF-κB antagonist QNZ and Bayl 1-7082, then HIF--1α and VEGF protein levels were determined with Western-blot,and expression levels of HIF-1α mRNA were quantified by RT-PCR method. RESULTS： EC signifi- cantly reduced the expression level of VEGF （P〈0.05）. The effect of clown regulation of VEGF expressions were also a- chieved in the three NHL cell lines transfected by pSuper HIF-1α/siRNA （P〈0. 05）. At the same time,inhibition of NF-κB significantly reduced HIF la protein and mRNA level as well as its related protein VEGF expression of human NHL cell in a dose dependent manner （P〈0.05）. CONCLUSION： The pathway of HIF-1α VEGF can be inhibited by In- hibition of NF-κB in human NHL cell in normoxic condition through down-regulation transfection of HIF-1α.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2012年第11期827-831,共5页
Chinese Journal of Cancer Prevention and Treatment
基金
辽宁省教育厅基金(2009A754)
